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人乳牙和恒牙牙髓肽能神经支配的免疫组织化学对比分析

Comparative immunohistochemical analysis of the peptidergic innervation of human primary and permanent tooth pulp.

作者信息

Rodd Helen D, Boissonade Fiona M

机构信息

School of Clinical Dentistry, University of Sheffield, Claremont Crescent, Sheffield S10 2TA, UK.

出版信息

Arch Oral Biol. 2002 May;47(5):375-85. doi: 10.1016/s0003-9969(02)00012-2.

Abstract

This immunohistochemical study sought to determine whether there are any differences in the peptidergic innervation of these pulps and whether dental caries is associated with changes in neuropeptide expression. Mandibular first permanent molars and second primary molars (n=120) were obtained from children requiring dental extractions under general anaesthesia. Extracted teeth were split longitudinally, placed in fixative, and categorized as intact, moderately carious or grossly carious. The coronal pulps were removed and 10-microm frozen sections were processed for indirect immunofluorescence. Double labelling employed combinations of the following antisera: (1) protein gene product 9.5, a general neuronal marker; (2) one of the neuropeptides calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin (GAL), enkephalin (ENK) and somatostatin (SOM). Image analysis was then used to determine the percentage area of immunostaining for each label within different anatomical regions of the coronal pulp. Sparse or absent immunoreactivity for GAL, ENK and SOM made analysis impossible. Analysis of CGRP, SP and VIP revealed significant interdentition differences, with their expression being significantly greater in permanent teeth, but this was not the case for NPY, with primary and permanent teeth demonstrating a similar amount of label for this peptide. Both dentitions showed significant increases in CGRP, SP, VIP and NPY expression with caries progression. These findings could have biological and clinical importance in connection with nociception, inflammation and healing.

摘要

这项免疫组化研究旨在确定这些牙髓的肽能神经支配是否存在差异,以及龋齿是否与神经肽表达的变化有关。从需要在全身麻醉下拔牙的儿童中获取下颌第一恒磨牙和第二乳磨牙(n = 120)。将拔除的牙齿纵向劈开,置于固定剂中,并分类为完整、中度龋或重度龋。去除冠髓,制作10微米厚的冰冻切片,进行间接免疫荧光处理。双重标记采用以下抗血清的组合:(1)蛋白基因产物9.5,一种通用的神经元标记物;(2)神经肽降钙素基因相关肽(CGRP)、P物质(SP)、血管活性肠肽(VIP)、神经肽Y(NPY)、甘丙肽(GAL)、脑啡肽(ENK)和生长抑素(SOM)中的一种。然后使用图像分析来确定冠髓不同解剖区域内每个标记物的免疫染色面积百分比。由于GAL、ENK和SOM的免疫反应性稀疏或缺失,无法进行分析。对CGRP、SP和VIP的分析显示出明显的牙间差异,它们在恒牙中的表达明显更高,但NPY并非如此,乳牙和恒牙中该肽的标记量相似。随着龋齿进展,两种牙列中CGRP、SP、VIP和NPY的表达均显著增加。这些发现可能在疼痛感受、炎症和愈合方面具有生物学和临床意义。

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